LOCUS       sequence-411019-        4346 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of R124A wgMDH with IPTG induction - Active 
            site mutant.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4346)
  AUTHORS   Cox B, Chit MM, Weaver T, Gietl C, Bailey J, Bell E, Banaszak L
  TITLE     Organelle and translocatable forms of glyoxysomal malate 
            dehydrogenase. The effect of the N-terminal presequence.
  JOURNAL   FEBS J. 2005 Feb;272(3):643-54. doi: 
            10.1111/j.1742-4658.2004.04475.x.
  PUBMED    15670147
REFERENCE   2  (bases 1 to 4346)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1111/j.1742-4658.2004.04475"; journalName: "FEBS J"; date: 
            "2005-02"; volume: "272"; issue: "3"; pages: "643-54"
COMMENT     Sequence Label: pQE60 R124A wgMDH
FEATURES             Location/Qualifiers
     source          1..4346
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    30..46
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     RBS             65..76
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             534..545
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     CDS             1049..1066
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      1089..1183
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(1293..1312)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1955..2041
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(2088..2110)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    2196..2336
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     2251..2270
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2351..2368)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2522..3110)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2602..2621)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3281..4072,4073..4141))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3904..3923
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4142..4246)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4314..4332
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ctcgagaaat cataaaaaat ttatttgctt tgtgagcgga taacaatttc acacagaatt
       61 cattaaagag gagaaattaa ctatggctaa aggcggagct cccgggttca aagtcgcaat
      121 acttggcgct gccggtggca ttggccagcc ccttgcgatg ttgatgaaga tgaatcctct
      181 ggtttctgtt ctacatctat atgatgtagt caatgcccct ggtgtcaccg ctgatattag
      241 ccacatggac acgggtgctg tggtgcgtgg attcttgggg cagcagcagc tggaggctgc
      301 gcttactggc atggatctta ttatagtccc tgcaggtgtt cctgcgaaac caggaatgac
      361 gagggatgat ctgttcaaaa taaacgcagg aattgtcaag actctgtgtg aagggattgc
      421 aaagtgttgt ccaagagcca ttgtcaacct gatcagtaat cctgtgaact ccaccgtgcc
      481 catcgcagct gaagttttca agaaggctgg aacttatgat ccaaagcgac ttctgggagt
      541 tacaatgctc gacgtagtca gagccaatac ctttgtggca gaagtattgg gtcttgatcc
      601 tcgggatgtt gatgttccag ttgttggcgg tcatgctggt gtaaccattt tgccccttct
      661 atctcaggtg aagcctccaa gttctttcac acaagaagag attagttacc tgactgatag
      721 gattcaaaat ggtggaacag aagttgtcga ggccaaagca ggagctggtt cagcaactct
      781 ctcaatggct tatgctgccg ttaagtttgc agatgcatgc ctcaggggct taagaggaga
      841 tgctggtgtc attgaatgcg cgtttgtgtc ttctcaggtg actgaacttc cattctttgc
      901 atcaaaagta cgacttggtc gcaatggtat cgaagaagta tactcccttg gcccgctaaa
      961 tgagtatgag aggattggat tggagaaagc gaagaaagag ttggcaggaa gcattgagaa
     1021 gggagtttcc ttcatcagaa gcagatctca tcaccatcac catcactaag cttaattagc
     1081 tgagcttgga ctcctgttga tagatccagt aatgacctca gaactccatc tggatttgtt
     1141 cagaacgctc ggttgccgcc gggcgttttt tattggtgag aatccaagct agcttggcga
     1201 gattttcagg agctaaggaa gctaaaatgg agaaaaaaat cactggatat accaccgttg
     1261 atatatccca atggcatcgt aaagaacatt ttgaggcatt tcagtcagtt gctcaatgta
     1321 cctataacca gaccgttcag ctggatatta cggccttttt aaagaccgta aagaaaaata
     1381 agcacaagtt ttatccggcc tttattcaca ttcttgcccg cctgatgaat gctcatccgg
     1441 aatttcgtat ggcaatgaaa gacggtgagc tggtgatatg ggatagtgtt cacccttgtt
     1501 acaccgtttt ccatgagcaa actgaaacgt tttcatcgct ctggagtgaa taccacgacg
     1561 atttccggca gtttctacac atatattcgc aagatgtggc gtgttacggt gaaaacctgg
     1621 cctatttccc taaagggttt attgagaata tgtttttcgt ctcagccaat ccctgggtga
     1681 gtttcaccag ttttgattta aacgtggcca atatggacaa cttcttcgcc cccgttttca
     1741 ccatgcatgg gcaaatatta tacgcaaggc gacaaggtgc tgatgccgct ggcgattcag
     1801 gttcatcatg ccgtttgtga tggcttccat gtcggcagaa tgcttaatga attacaacag
     1861 tactgcgatg agtggcaggg cggggcgtaa tttttttaag gcagttattg gtgcccttaa
     1921 acgcctgggg taatgactct ctagcttgag gcatcaaata aaacgaaagg ctcagtcgaa
     1981 agactgggcc tttcgtttta tctgttgttt gtcggtgaac gctctcctga gtaggacaaa
     2041 tccgccctct agagctgcct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg
     2101 cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt
     2161 cagggcgcgt cagcgggtgt tggcgggtgt cggggcgcag ccatgaccca gtcacgtagc
     2221 gatagcggag tgtatactgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
     2281 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgct
     2341 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     2401 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     2461 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     2521 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     2581 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     2641 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     2701 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     2761 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     2821 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     2881 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     2941 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta
     3001 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     3061 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     3121 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     3181 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     3241 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     3301 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     3361 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     3421 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     3481 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     3541 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     3601 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     3661 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     3721 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     3781 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     3841 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     3901 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     3961 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     4021 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     4081 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     4141 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     4201 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     4261 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc
     4321 gtatcacgag gccctttcgt cttcac
//
